增脂耶氏菌奇链脂肪酸的工程前体池

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Metabolic Engineering Communications Pub Date : 2021-06-01 DOI:10.1016/j.mec.2020.e00158
Young-Kyoung Park , Florence Bordes , Fabien Letisse , Jean-Marc Nicaud
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引用次数: 16

摘要

随着环境和能源的日益关注,微生物生产的脂质是化石资源的有前途的替代品之一。奇链脂肪酸(OCFA)是一种特殊的脂类,由于其在医疗、制药和化学工业中的广泛应用,近年来作为微生物生产的目标化合物受到了广泛的关注。在这项研究中,我们的目的是增加三碳链(丙炔- coa)和五碳链(β-酮戊酰- coa)的前体池,用于在脂性耶氏菌中生产OCFAs。我们对不同的丙酸激活酶进行了评价,结果表明,过表达丙酰coa转移酶基因的富菌OCFAs的积累量比对照菌株增加了3.8倍,表明丙酸激活是OCFAs合成的限制步骤。结果表明,为了恢复生长和提高总脂质中OCFA的含量,补充乙酸是必要的,这表明乙酰辅酶a和丙酰辅酶a前体之间的平衡对OCFA的积累至关重要。为了改善β-酮戊酰辅酶a库,进一步提高OCFA产量,我们在产菌中共表达了编码β-酮硫酶的bktB, OCFA产量比对照提高了33%。结合菌株工程和优化C/N比,使OCFA产量达到1.87 g/L,占总脂质的62%,是迄今为止在酵母中报道的最高重组OCFA滴度。该研究为微生物生产OCFAs及其衍生物提供了强有力的基础,具有广泛的应用潜力。
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Engineering precursor pools for increasing production of odd-chain fatty acids in Yarrowia lipolytica

Microbial production of lipids is one of the promising alternatives to fossil resources with increasing environmental and energy concern. Odd-chain fatty acids (OCFA), a type of unusual lipids, are recently gaining a lot of interest as target compounds in microbial production due to their diverse applications in the medical, pharmaceutical, and chemical industries. In this study, we aimed to enhance the pool of precursors with three-carbon chain (propionyl-CoA) and five-carbon chain (β-ketovaleryl-CoA) for the production of OCFAs in Yarrowia lipolytica. We evaluated different propionate-activating enzymes and the overexpression of propionyl-CoA transferase gene from Ralstonia eutropha increased the accumulation of OCFAs by 3.8 times over control strain, indicating propionate activation is the limiting step of OCFAs synthesis. It was shown that acetate supplement was necessary to restore growth and to produce a higher OCFA contents in total lipids, suggesting the balance of the precursors between acetyl-CoA and propionyl-CoA is crucial for OCFA accumulation. To improve β-ketovaleryl-CoA pools for further increase of OCFA production, we co-expressed the bktB encoding β-ketothiolase in the producing strain, and the OCFA production was increased by 33% compared to control. Combining strain engineering and the optimization of the C/N ratio promoted the OCFA production up to 1.87 ​g/L representing 62% of total lipids, the highest recombinant OCFAs titer reported in yeast, up to date. This study provides a strong basis for the microbial production of OCFAs and its derivatives having high potentials in a wide range of applications.

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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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